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[DNA细胞计量学中的标准化和质量保证问题]

[Problems concerning standardization and quality assurance in DNA cytometry].

作者信息

Aubele M, Rodenacker K

机构信息

GSF-Forschungszentrum für Umwelt und Gesundheit GmbH, Institut für Pathologie, Oberschleissheim, Deutschland.

出版信息

Zentralbl Pathol. 1994 Feb;139(6):437-41.

PMID:7512822
Abstract

Rat liver imprints were treated with five different fixation techniques. Chicken erythrocytes stored over an extended period in the refrigerator were dropped on each slide before Feulgen staining. By means of an image analysis system chicken erythrocytes, rat leukocytes and hepatocytes were measured and the integrated optical density (IOD) was calculated for each nucleus. The coefficient of variation (CV) of IOD was about 15% for chicken erythrocytes. Leukocytes showed a CV of up to 10%. The CV was lower for hepatocytes than for leukocytes, and lowest for air-dried hepatocytes. The standard error of the mean (SEM) did not show remarkable differences between the fixation groups. For hepatocytes it was in general less than 1% of the respective mean value. The hepatocytes showed a linear staining except the wet formalin-fixed cells. This holds also for the wet formalin fixed leukocytes which showed only 85% of the IOD of 2c hepatocytes. The ratios of erythrocytes to 2c hepatocytes varied between 0.33 and 0.35. The IOD ratios of standard cells to 2c hepatocytes from single specimens showed remarkable differences from the above mentioned ratios of the pooled slides of a fixation group. The use of external standard cells was proved to be problematic, especially because of their variation in staining intensity independent of the staining intensity of hepatocytes.

摘要

大鼠肝脏印片采用五种不同的固定技术处理。在福尔根染色前,将长期保存在冰箱中的鸡红细胞滴加到每张载玻片上。借助图像分析系统,对鸡红细胞、大鼠白细胞和肝细胞进行测量,并计算每个细胞核的积分光密度(IOD)。鸡红细胞的IOD变异系数(CV)约为15%。白细胞的CV高达10%。肝细胞的CV低于白细胞,风干的肝细胞CV最低。固定组之间的平均标准误差(SEM)没有显著差异。对于肝细胞,一般低于各自平均值的1%。除湿福尔马林固定的细胞外,肝细胞呈现线性染色。湿福尔马林固定的白细胞也是如此,其IOD仅为2c肝细胞的85%。红细胞与2c肝细胞的比例在0.33至0.35之间。单个标本中标准细胞与2c肝细胞的IOD比值与固定组合并载玻片的上述比值存在显著差异。事实证明,使用外部标准细胞存在问题,特别是因为它们的染色强度变化与肝细胞的染色强度无关。

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